scholarly journals EFFECT OF INFLUENZA VIRUS ON CILIA AND EPITHELIAL CELLS IN THE BRONCHI OF MICE

1952 ◽  
Vol 95 (2) ◽  
pp. 173-189 ◽  
Author(s):  
Carl G. Harford ◽  
Alice Hamlin
Keyword(s):  
Influenza Virus ◽  
Viral Infection ◽  
Cellular Proliferation ◽  
Pneumococcal Infection ◽  
Bronchial Epithelium ◽  
Ciliary Activity ◽  
Polymorphonuclear Leucocytes ◽  
Fixed Tissue ◽  
Staining Methods ◽  
Ciliary Beat

In order to determine the effect of infection with influenza virus on bronchial cilia of the mouse, ciliary beat has been visualized directly by microscopic examination of the bronchi in slices of fresh lung. Cilia have been shown also in sections of fixed tissue by the use of special silver staining methods. The results have shown persistence of the cilia in spite of severe viral infection and indicate that the lowered resistance to secondary pneumococcal infection which occurs in influenzal pneumonia of the mouse is not due to interference with the ciliary mechanism. By a process of exclusion, the findings give further support to the theory that lowered resistance to pneumococcal infection in influenzal pneumonia is due to edema fluid in the viral lesion furnishing a culture medium for inhaled pneumococci. A widely used method for evaluation of ciliary activity on respiratory epithelia has been the microscopic observation of wave-like movements in reflected light. This activity was observed readily in the bronchi of mice but evidence was obtained showing that at this site it was due to something other than ciliary beat. Further histopathologic observations were made in order to define the lesion of the bronchial epithelium that would permit sparing of ciliated cells. In addition to usual techniques, mice were injected with colchicine for estimation of the rate of cellular proliferation and were exposed to a large dose of roentgen rays to eliminate polymorphonuclear leucocytes. Stains for mucous and for mitochondria were done also. The evidence obtained favors the theory that the viral infection does not destroy any of the cells of the bronchial epithelium. Inclusion bodies were found in the cytoplasm, making it seem likely instead that viral particles grow in colony-like aggregations and that liberation of virus into the lumen takes place not only by simple extrusion of inclusions but also by detachment of inclusion-laden globular portions of the cytoplasm.

Enzyme Histochemistry of Bronchial Epithelium and Alveolar Cells in the Early Stages of Influenza-Virus Pneumonia of Mice

Nature ◽  
1962 ◽  
Vol 193 (4813) ◽  
pp. 348-349 ◽  
Author(s):  
J. F. Ph. HERS ◽  
R. G. J. WILLIGHAGEN ◽  
D. A. J. TYRRELL ◽  
LIDY van der KUIP ◽  
MARIA A. C. HOS
Keyword(s):  
Influenza Virus ◽  
Enzyme Histochemistry ◽  
Bronchial Epithelium ◽  
Alveolar Cells ◽  
Early Stages ◽  
Virus Pneumonia

Inhibition of ciliary activity in organ cultures of ferret trachea in reference to genetic and biological characters of influenza virus strains

1982 ◽  
Vol 28 (7) ◽  
pp. 809-814 ◽  
Author(s):  
P. Diaz-Rodriguez ◽  
A. Boudreault
Keyword(s):  
Influenza Virus ◽  
Influenza Viruses ◽  
Influenza Vaccines ◽  
Ciliary Activity ◽  
Organ Cultures ◽  
Preliminary Screening ◽  
Cold Adapted ◽  
Activity Inhibition ◽  
Biological Characters ◽  
Virus Strains

As reported previously, attenuated stable inhibitor-resistant influenza viruses can be screened by a 50% ciliary activity inhibition test in ferret tracheal organ cultures. This test was further applied to 5 attenuated cold-adapted influenza strains and to 11 strains with known a percentage of RNA–RNA hybridization with the parental A/PR/8/34 (H0N1) virus strain. Again, with one exception, attenuated strains could be clearly differentiated from virulent ones. It was concluded that virulence of influenza strains for man can be detected using this test regardless of the techniques used to prepare attenuated variants. A preliminary screening of attenuated candidates for live influenza vaccines can be achieved with confidence on ferret tracheal organ cultures.

scholarly journals Effect of the preparation Ingavirin® (imidazolyl ethanamide pentandioic acid) on the interferon status of cells under conditions of viral infection

2016 ◽  
Vol 21 (4) ◽  
pp. 196-205
Author(s):  
Thomas Aschacher ◽  
Artem Krokhin ◽  
Irina Kuznetsova ◽  
Johannes Langle ◽  
Vladimir Nebolsin ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Viral Infection ◽  
Viral Infections ◽  
Antiviral Drug ◽  
Effector Proteins ◽  
Theoretical Ground ◽  
Ns1 Protein ◽  
Interferon Inducer ◽  
Infected Cells ◽  
Interferon System

Ingavirin® (imidazolyl ethanamide pentandioic acid) is an original antiviral drug, which is used in Russia for treatment and profilaxis of influenza and other acute viral infections. We confirmed that imidazolyl ethanamide pentandioic acid (IEPA), not being interferon inducer itself, enhances synthesis of both interferon-a/fi receptors (IFNAR) to interferone and cell sensitivity to interferone signalling, which was suppressed by NS1 protein - pathogen factor of influenza virus. IEPA is able to promote antiviral effector proteins PKR and MxA in infected cells, in opposition to interferon system suppression by influenza virus. Theoretical ground of clinical efficacy of Ingavirine® could be confirmed by obtained data of influence to innate immune system during viral infection.

scholarly journals Brugada-Like Electrocardiographic Changes during Influenza Infection

2003 ◽  
Vol 31 (3) ◽  
pp. 244-246 ◽  
Author(s):  
K Kusaka ◽  
J Yamakawa ◽  
K Kawaura ◽  
T Itoh ◽  
T Takahashi ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Virus Infection ◽  
Viral Infection ◽  
Brugada Syndrome ◽  
Influenza Infection ◽  
Influenza Virus Infection ◽  
Ecg Changes ◽  
Early Repolarization ◽  
Electrocardiographic Changes

We describe a 32-year-old man with electrocardiographic (ECG) changes consistent with Brugada syndrome and influenza virus infection. The ECG pattern changed after 1 week to one of early repolarization in V1 and V2. This case suggests an association between Brugada syndrome and viral infection.

Evaluation of cytokine production by J774.G8 macrophages after treatment with biotherapics

2021 ◽  
Vol 10 (36) ◽  
pp. 167-169
Author(s):  
Camila Siqueira ◽  
Diogo Kuczera ◽  
Eneida Da Lozzo ◽  
Dorly Buchi ◽  
José Nelson Couceiro ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Viral Infection ◽  
Influenza A ◽  
Control Groups ◽  
Viral Particles ◽  
Significant Difference ◽  
Infected Cells ◽  
H3n2 Influenza ◽  
Aseptic Conditions ◽  
H3n2 Influenza Virus

Introduction: Strains of macrophages, such as murine J774.G8 macrophages, are susceptible to influenza A infection [1]. One of the responses to viral infection involves the production of various types of immunostimulatory cytokines by infected cells [2]. Methods: In the present study, the macrophage strain J774.G8, maintained in RPMI medium, was submitted to treatment with 10% V/V of two different biotherapics prepared from influenza H3N2, both at 30x. Additionally, two control groups were analyzed: macrophages stimulated with water 30x and macrophages without any treatment. Biotherapics were prepared from intact H3N2 influenza virus and H3N2 inactivated by alcohol 70%. The compounding of both biotherapics followed this procedure: one part of viral particles was diluted in 9 parts of sterile distilled water. The 1:10 sample was submitted to 100 mechanical succussions using Autic® Brazilian machine, originating the first dilution, named decimal (1x). 1 ml of this solution was diluted in 9 ml of solvent and was submitted to 100 succussions, generating biotherapic 2x. This procedure was successively repeated, according to Brazilian Homeopathic Pharmacopoeia, to obtain the biotherapic 30x. By the same technique, water vehicle was prepared in the potency of 30x to be used as control. All samples were prepared under sterile and aseptic conditions, using laminar flow cabinet, class II, and were stored in the refrigerator (8ºC), to avoid microbiological contamination. J774.G8 macrophages were stimulated for 2 days, in a total of six stimuli. Immediately before infection with 25 µl of H3N2 influenza virus, the supernatants were collected and frozen at -20 ºC for later analysis. Next, 24 hours after the virus infection, the supernatants were aliquoted and frozen under the same conditions. Three independent experiments were done in triplicate. Analysis of supernatants was performed by flow cytometry using the Mouse Inflammation Kit. The cytokines detected in this experiment were IL-10, IL 12, TNF-α and MCP1. Results: In all cases, there were no significant differences compared to control groups. However, the production of TNF-α detected in macrophages treated by intact and inactivated biotherapics presented a tendency to increase after infection. In fact, similar results were previously detected in other experiments conducted only with the intact biotherapic [3]. The release of the cytokine MCP1 in all experimental situations presented a tendency to decrease after the viral infection when compared to untreated macrophages. No statistically significant difference was detected in the production of IL 12 and IL 10. These experiments will be repeated to confirm the data obtained.

Combined Effect of Influenza Virus Infection and Urethan Treatment on the Incidence of Lung: Tumors in Mice.

1965 ◽  
Vol 51 (6) ◽  
pp. 401-417 ◽  
Author(s):  
Anna Maria Casazza ◽  
Marcello Gaetani ◽  
Mario Ghione ◽  
Enrico Turolla
Keyword(s):  
Influenza Virus ◽  
Virus Infection ◽  
Joint Action ◽  
Influenza A ◽  
Influenza Virus Infection ◽  
Alveolar Epithelium ◽  
Bronchial Epithelium ◽  
Lung Tumors ◽  
Pulmonary Tissue ◽  
Two Agents

Swiss mice were intranasally infected with influenza A2 virus and treated with urethan in order to detect whether the joint action of the two agents would enhance the development of lung tumors. The average number per mouse of the typical lesions induced by the two treatments together with their location, their histological and histochemical characteristics and the percentage of death in the different groups of animals were recorded. Results indicated that 51.7 % of the mice infected with influenza virus and treated with urethan had both bronchial dysplastic lesions due to influenza virus, and tumors induced by urethan. In this group the number of tumors was smaller than in the mice treated with the carcinogen only and no invasive pulmonary carcinomas were observed. The dysplastic lesions caused by influenza A2 virus as well as the lung adenomas induced by urethan maintained their typical histological and histochemical characteristics even when occurring in a close position. The failure of urethan to enhance the induction of lung tumors in mice exposed to influenzal infection might be ascribed to the different primary sites of response of the pulmonary tissue to the two agents, i.e. the bronchial epithelium for influenza virus and the alveolar epithelium for urethan.

HSP27 elevated in mild allergic inflammation protects airway epithelium from H2SO4 effects

1997 ◽  
Vol 273 (2) ◽  
pp. L401-L409 ◽  
Author(s):  
A. T. Hastie ◽  
K. B. Everts ◽  
J. Zangrilli ◽  
J. R. Shaver ◽  
M. B. Pollice ◽  
...  
Keyword(s):  
Airway Epithelium ◽  
Stress Proteins ◽  
Stress Protein ◽  
Allergic Inflammation ◽  
Allergen Challenge ◽  
Bronchial Epithelium ◽  
Ciliary Activity ◽  
Normal Epithelium ◽  
Adverse Conditions ◽  
Before And After

Inflammation in allergic individuals is hypothesized to elevate stress proteins [heat shock proteins (HSP)] in airway epithelium, which may protect cells from further adverse conditions. Allergic, either asthmatic or not, and normal volunteers participated in a 2-day segmental allergen challenge bronchoscopic procedure. Bronchial epithelium was obtained before and after challenge. Epithelium was exposed to medium with H2SO4 (pH5), returned to medium at pH 7.4, and finally harvested for Western blotting with anti-27-kDa HSP (HSP27) antibody. Prechallenge epithelium of all subjects had significantly inhibited ciliary function by H2SO4 (pH 5) conditions (P < 0.001); only epithelium of normals recovered (P = 0.02). Allergic subjects with mild inflammation (< 50 micrograms/ml increase in albumin in bronchoalveolar lavage) had significantly increased HSP27 postchallenge (P = 0.01) and little ciliary dysfunction at pH 5, whereas subjects with severe inflammation (> 50 micrograms/ml increase in albumin) had little change in HSP27 and significant ciliary inhibition (P = 0.02). Normal epithelium had similar trends in HSP27 and equivalent inhibition of ciliary activity at pH 5 before and after allergen challenge. These data indicate that mild inflammation to allergen elevates HSP27 stress protein levels, thereby potentially protecting epithelial function from additional adverse conditions.

Honorable Mention — Student Research Award 1995: Signal Transduction Mechanisms in Substance P-Mediated Ciliostimulation

1995 ◽  
Vol 113 (5) ◽  
pp. 582-588 ◽  
Author(s):  
Rodney J. Schlosser ◽  
Judith M. Czaja ◽  
Thomas V. McCaffrey
Keyword(s):  
Nitric Oxide ◽  
Substance P ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Intermediate Step ◽  
Nitric Oxide Production ◽  
Oxide Production ◽  
Vitro Effect ◽  
Ciliary Beat

Substance P is a neuropeptide released by afferent neurons in the respiratory tract during inflammatory reactions. It produces effects on blood vessels, bronchial smooth muscle, nasal glands, and respiratory cilia. We studied the in vitro effect of substance P on the ciliary beat frequency of human adenoid explants and its mechanism of action. Substance P was added to cultured adenoid at concentrations of 10−10, 10−8, 10−6, and 10−4 mol/L. Ciliary beat frequency was determined with phase-contrast microscopy and microphotometry. Substance P increased ciliary beat frequency a maximum of 11.9% ± 3.8% ( p < 0.01). Diclofenac (10−6 mol/L) significantly blocked the ciliostimulatory effects of SP ( p < 0.022), indicating that prostaglandin synthesis is an intermediate step in the action of substance P on ciliary beat frequency. The L-arginine analogs, NG-nitro-L-arginine methyl ester and NG-monomethyl-L-arginine, inhibit nitric oxide synthesis from L-arginine. L-Arginine analogs (10−4 to 10−2 mol/L) inhibited the effect of substance P ( p < 0.02 at the higher concentration). This inhibition was reversed by adding L-arginine, demonstrating that nitric oxide production is a required step in substance P-induced ciliostimulation. Substance P stimulates ciliary activity in human nasal mucosa as a result of secondary production and release of endogenous prostaglandins and nitric oxide. It is likely that inflammatory disease processes that stimulate release of substance P and subsequent prostaglandin and nitric oxide production modify mucociliary transport. Pharmacologic modification of substance P and its second messengers may eventually permit regulation of this important defense mechanism and control of neurogenic inflammation.

111 Inhibition of Avian Influenza Virus by Blocking Specific Sialyltransferases

2018 ◽  
Vol 30 (1) ◽  
pp. 195
Author(s):  
E. N. Antonova ◽  
O. V. Glazova ◽  
A. V. Gaponova ◽  
N. A. Volkova ◽  
P. Y. Volchkov
Keyword(s):  
Flow Cytometry ◽  
Influenza Virus ◽  
Sialic Acid ◽  
Avian Influenza ◽  
Viral Infection ◽  
Cell Line ◽  
Influenza A ◽  
Binding Assay ◽  
Fluorescein Isothiocyanate ◽  
Sialic Acids

It is known that avian influenza penetrates into the host cell by binding with sialic acids, the terminal residues of oligosaccharides. Avian influenza A virus preferably recognises α(2,3)-linked sialic acid residues as a receptor for penetration whereas human influenza A virus preferably binds with α(2,6)-linked sialic acids. Prevention of transfer of sialic acids to sugar bond or removal of it could be a defensive strategy against viral infection. There are 6 known sialyltransferases (ST3Gal1-6) that transfer α(2,3)-linked sialic acid residues to sugar branches. Most avian influenza virus isolates bind strongly to a sugar chain containing Neu5Aca(2,3) residues. In our study, we have shown that knockout of sialyltransferases leads to inhibition of viral infection. To find the expressed sialyltransferases in respiratory and digestive tracts, we used RT-qPCR. Tissue samples were taken from 3 chickens of Haisex white cross. Expression of mRNA was measured by RT-qPCR in 3 repeats and serial dilutions. Data analysis was carried out using the 2−ΔΔCt method. The amount of total RNA was normalised using GAPDH mRNA. For CRISPR/Cas9 targeting sialyltransferases, 3 guide RNAs for each gene were designed. We confirmed knockout (KO) of ST3GAL1 and ST3GAL6 by T7E assay. To estimate sialylation level on the cell surface, we performed a lectin-binding assay. For the assay, cells were incubated with fluorescein isothiocyanate (FITC)-labelled Maackia amurensis lectins and then subjected to flow-cytometry analysis to quantify the percentage of α(2,3)-sialylated cells in DF1 knockout (KO) v. DF1 wild type (wt) cell line. To estimate resistance to viral infection, a hemagglutinin binding assay was done, using fluorescein isothiocyanate (FITC)-labelled HA1 from H5N1 (A/Vietnam/1203/2004). To quantify the percentage of agglutinated HA1 molecules, DF1 KO and DF1 wt cells were analysed by flow cytometry. We found that mainly ST3GAL4 and ST3GAL5 are expressed in the chicken intestine (3-fold and 20-fold less compared with GAPDH level, respectively; other STs were not detected), and mainly ST3GAL1 and ST3GAL6 are expressed in the chicken respiratory tract (5-fold and 1.2-fold more compared with GAPDH level respectively; other STs were not detected). The expression profile of α(2,3)-sialyltransferases in the DF1 chicken cell line showed the noticeable expression of ST3GAL1 and ST3GAL6 compared with others as has been shown for the respiratory tract (500- and 1000-fold less compared with GAPDH respectively; other STs were not detected). In this study, we adopted the CRISPR/Cas9 system to knock out ST3GAL1 and ST3GAL6 genes in the chicken DF1 cell line. We confirmed that knockout of the genes leads to extinction of α(2,3)-sialic residues from the cell surface (7% v. 100% for DF1 KO v. DF1 wt cell line). Finally, we showed that knockout of sialyltransferases in the DF1 cells increases resistance against influenza A infection (16% v. 100% for DF1 KO v. DF1 wt cell line). Thus, creation of transgenic poultry with tissue-specific knockout of the α(2,3) sialyltransferases might protect domestic birds against influenza virus and block possible transfer of avian flu to human population.

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